Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Dec 15;7:136. doi: 10.1038/s41698-023-00486-6

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 4 — a HEK293T cells were transfected with MYC-tagged SMURF2 and/or GST-tagged SHP2. After 48 h, cells were lysed and immunoprecipitated with GST antibody. Immunoprecipitated lysates and whole cell extracts were probed with the indicated antibodies. b HEK293T cells were transfected with MYC-tagged SMURF2 and/or GST-tagged SHP2. After 48 h, cells were lysed and immunoprecipitated with MYC antibody. Immunoprecipitated lysates and whole cell extracts were probed with the indicated antibodies. c HEK293T cells transduced with SHP2 or relevant controls along with MYC-tagged SMURF2 and FLAG-tagged c-SRC. Lysates were immunoprecipitated with anti-MYC affinity resin. Immunoprecipitated lysates and whole cell extracts were probed with the indicated antibodies. pY signifies tyrosine phosphorylation. d HEK293T cells transduced with siRNA targeting SHP2 or relevant controls along with MYC-tagged SMURF2 and FLAG-tagged c-SRC. Lysates were immunoprecipitated with anti-MYC affinity resin. Immunoprecipitated lysates and whole cell extracts were probed with the indicated antibodies. pY signifies tyrosine phosphorylation. e HEK293T cells transduced with SHP2, FLAG-tagged c-SRC, and MYC-tagged WT SMURF2, or SMURF2 mutant (SMURF2 Y314F/Y434F, (SMURF2 FF)). Lysates were immunoprecipitated with anti-MYC affinity resin. Immunoprecipitated lysates and whole cell extracts were probed with the indicated antibodies. Y314 phosphorylation was determined with a SMURF2 specific Y314 phospho-antibody. f HEK293T cells transduced with SHP2, FLAG-tagged c-SRC, and MYC-tagged WT SMURF2, or SMURF2 mutant (SMURF2 Y314F/Y434F, (SMURF2 FF)). Lysates were immunoprecipitated with anti-MYC affinity resin. Immunoprecipitated lysates and whole cell extracts were probed with the indicated antibodies. Y434 phosphorylation was determined with a SMURF2 specific Y434 phospho-antibody. g HEK293T cells were transfected as indicated with MYC-tagged SMURF2, Flag-tagged SMAD7, and/or GST-tagged SHP2. After 48 h cells were lysed and immunoprecipitated with anti-FLAG affinity resin. Immunoprecipitated lysates and whole cell extracts were probed with the indicated antibodies. h HEK293T cells transfected with wild-type MYC-SMURF2, HA-tagged ubiquitin and/or GST-SHP2. Following immunoprecipitation of MYC-SMURF2, lysates were resolved by SDS-PAGE and probed with indicated antibodies. i TGFβ responsive luciferase (CAGA luciferase) of H358 cells stimulated overnight with TGFβ (100 pM) or SHP099 (10 μM) or in combination with or without ectopic expression of SMURF2 EE or SMURF2 FF. Lysates were collected and luciferase measured by a luminometer. Error bars represent S.D. of triplicates. Experiments are representative of three independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001 as determined by Student’s t-test.